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Apoptosis

, Volume 16, Issue 4, pp 347–358 | Cite as

Panaxydol induces apoptosis through an increased intracellular calcium level, activation of JNK and p38 MAPK and NADPH oxidase-dependent generation of reactive oxygen species

  • Joo Young Kim
  • Su-Jin Yu
  • Hyun Ju Oh
  • Ji Young Lee
  • Yongjin Kim
  • Jeongwon Sohn
Original Paper

Abstract

Panaxydol, a polyacetylenic compound derived from Panax ginseng roots, has been shown to inhibit the growth of cancer cells. In this study, we demonstrated that panaxydol induced apoptosis preferentially in transformed cells with a minimal effect on non-transformed cells. Furthermore, panaxydol was shown to induce apoptosis through an increase in intracellular Ca2+ concentration ([Ca2+]i), activation of JNK and p38 MAPK, and generation of reactive oxygen species (ROS) initially by NADPH oxidase and then by mitochondria. Panaxydol-induced apoptosis was caspase-dependent and occurred through a mitochondrial pathway. ROS generation by NADPH oxidase was critical for panaxydol-induced apoptosis. Mitochondrial ROS production was also required, however, it appeared to be secondary to the ROS generation by NADPH oxidase. Activation of NADPH oxidase was demonstrated by the membrane translocation of regulatory p47phox and p67phox subunits and shown to be necessary for ROS generation by panaxydol treatment. Panaxydol triggered a rapid and sustained increase of [Ca2+]i, which resulted in activation of JNK and p38 MAPK. JNK and p38 MAPK play a key role in activation of NADPH oxidase, since inhibition of their expression or activity abrogated membrane translocation of p47phox and p67phox subunits and ROS generation. In summary, these data indicate that panaxydol induces apoptosis preferentially in cancer cells, and the signaling mechanisms involve a [Ca2+]i increase, JNK and p38 MAPK activation, and ROS generation through NADPH oxidase and mitochondria.

Keywords

Panaxydol Apoptosis Reactive oxygen species NADPH oxidase Calcium MAP kinase 

Notes

Acknowledgments

This work was supported by a KOSEF grant (2009-0080582) and the National Research Foundation of Korea (NRF) grant funded by the Korean Ministry of Education, Science and Technology (R0809661).

Supplementary material

10495_2010_567_MOESM1_ESM.docx (490 kb)
Supplementary material 1 (DOCX 489 kb)

References

  1. 1.
    Moon J, Yu SJ, Kim HS, Sohn J (2000) Induction of G(1) cell cycle arrest and p27(KIP1) increase by panaxydol isolated from Panax ginseng. Biochem Pharmacol 59:1109–1116PubMedCrossRefGoogle Scholar
  2. 2.
    Hai J, Lin Q, Lu Y, Zhang H, Yi J (2007) Induction of apoptosis in rat C6 glioma cells by panaxydol. Cell Biol Int 31:711–715PubMedCrossRefGoogle Scholar
  3. 3.
    Ryter SW, Kim HP, Hoetzel A et al (2007) Mechanisms of cell death in oxidative stress. Antioxid Redox Signal 9:49–89PubMedCrossRefGoogle Scholar
  4. 4.
    Buttke TM, Sandstrom PA (1994) Oxidative stress as a mediator of apoptosis. Immunol Today 15:7–10PubMedCrossRefGoogle Scholar
  5. 5.
    Jacobson MD (1996) Reactive oxygen species and programmed cell death. Trends Biochem Sci 21:83–86PubMedGoogle Scholar
  6. 6.
    Miyajima A, Nakashima J, Yoshioka K (1997) Role of reactive oxygen species in cis-dichlorodiammineplatium-induced cytotoxicity on bladder cancer cells. Br J Cancer 76:206–210CrossRefGoogle Scholar
  7. 7.
    Zhou Y, Hileman EO, Plunkett W, Keating MJ, Huang P (2003) Free radical stress in chronic lymphocytic leukemia cells and its role in cellular sensitivity to ROS-generating anticancer agents. Blood 101:4098–4104PubMedCrossRefGoogle Scholar
  8. 8.
    Pelicano H, Feng L, Zhou Y et al (2003) Inhibition of mitochondrial respiration: a novel strategy to enhance drug-induced apoptosis in human leukemia cells by a reactive oxygen species-mediated mechanism. J Biol Chem 278:37832–37839PubMedCrossRefGoogle Scholar
  9. 9.
    Schulze-Osthoff K, Bakker AC, Vanhaesebroeck B, Beyaert R, Jacob WA, Fiers W (1992) Cytotoxic activity of tumor necrosis factor is mediated by early damage of mitochondrial functions. Evidence for the involvement of mitochondrial radical generation. J Biol Chem 267:5317–5323PubMedGoogle Scholar
  10. 10.
    Quillet-Mary A, Jaffrezou JP, Mansat V, Bordier C, Naval J, Laurent G (1997) Implication of mitochondrial hydrogen peroxide generation in ceramide-induced apoptosis. J Biol Chem 272:21388–21395PubMedCrossRefGoogle Scholar
  11. 11.
    Fleury C, Mignotte B, Vayssiere JL (2002) Mitochondrial reactive oxygen species in cell death signaling. Biochimie 84:131–141PubMedCrossRefGoogle Scholar
  12. 12.
    Ott M, Gogvadze V, Orrenius S, Zhivotovsky B (2007) Mitochondria, oxidative stress and cell death. Apoptosis 12:913–922PubMedCrossRefGoogle Scholar
  13. 13.
    Hiraoka W, Vazquez N, Nieves-Neira W, Chanock SJ, Pommier Y (1998) Role of oxygen radicals generated by NADPH oxidase in apoptosis induced in human leukemia cells. J Clin Invest 102:1961–1968PubMedCrossRefGoogle Scholar
  14. 14.
    Qin F, Patel R, Yan C, Liu W (2006) NADPH oxidase is involved in angiotensin II-induced apoptosis in H9C2 cardiac muscle cells: effects of apocynin. Free Radic Biol Med 40:236–246PubMedCrossRefGoogle Scholar
  15. 15.
    Brennan AM, Suh SW, Won SJ et al (2009) NADPH oxidase is the primary source of superoxide induced by NMDA receptor activation. Nat Neurosci 12:857–863PubMedCrossRefGoogle Scholar
  16. 16.
    Nicotera P, Orrenius S (1998) The role of calcium in apoptosis. Cell Calcium 23:173–180PubMedCrossRefGoogle Scholar
  17. 17.
    Granfeldt D, Samuelsson M, Karlsson A (2002) Capacitative Ca2+ influx and activation of the neutrophil respiratory burst. Different regulation of plasma membrane- and granule-localized NADPH-oxidase. J Leukoc Biol 71:611–617PubMedGoogle Scholar
  18. 18.
    Wang G, Anrather J, Glass MJ et al (2006) Nox2, Ca2+, and protein kinase C play a role in angiotensin II-induced free radical production in nucleus tractus solitaries. Hypertension 48:482–489PubMedCrossRefGoogle Scholar
  19. 19.
    Klein-Szanto AJ, Iizasa T, Momiki S et al (1992) A tobacco-specific N-nitrosamine or cigarette smoke condensate causes neoplastic transformation of xenotransplanted human bronchial epithelial cells. Proc Natl Acad Sci USA 89:6693–6697PubMedCrossRefGoogle Scholar
  20. 20.
    Kim JE, Koo KH, Kim YH, Sohn J, Park YG (2008) Identification of potential lung cancer biomarkers using an in vitro carcinogenesis model. Exp Mol Med 40:709–720PubMedCrossRefGoogle Scholar
  21. 21.
    Kelso GF, Porteous CM, Coulter CV et al (2001) Selective targeting of a redox-active ubiquinone to mitochondria within cells: antioxidant and antiapoptotic properties. J Biol Chem 276:4588–4596PubMedCrossRefGoogle Scholar
  22. 22.
    Lee JY, Yu SJ, Park YG, Kim J, Sohn J (2007) Glycogen synthase kinase 3beta phosphorylates p21WAF1/CIP1 for proteasomal degradation after UV irradiation. Mol Cell Biol 27:3187–3198PubMedCrossRefGoogle Scholar
  23. 23.
    Matsuzawa A, Ichijo H (2008) Redox control of cell fate by MAP kinase: physiological roles of ASK1-MAP kinase pathway in stress signaling. Biochim Biophys Acta 1780:1325–1336PubMedGoogle Scholar
  24. 24.
    Kuppusamy P, Li H, Ilangovan G et al (2002) Noninvasive imaging of tumor redox status and its modification by tissue glutathione levels. Cancer Res 62:307–312PubMedGoogle Scholar
  25. 25.
    Jacobson MD, Raff MC (1995) Programmed cell death and Bcl-2 protection in very low oxygen. Nature 374:814–816PubMedCrossRefGoogle Scholar
  26. 26.
    Scorrano L, Oakes SA, Opferman JT et al (2003) BAX and BAK regulation of endoplasmic reticulum Ca2+: a control point for apoptosis. Science 300:135–139PubMedCrossRefGoogle Scholar
  27. 27.
    Yu JH, Lim JW, Kim KH, Morio T, Kim H (2005) NADPH oxidase and apoptosis in cerulein-stimulated pancreatic acinar AR42J cells. Free Radic Biol Med 39:590–602PubMedCrossRefGoogle Scholar
  28. 28.
    Gandhi S, Wood-Kaczmar A, Yao Z et al (2009) PINK1-associated Parkinson’s disease is caused by neuronal vulnerability to calcium-induced cell death. Mol Cell 33:627–638PubMedCrossRefGoogle Scholar
  29. 29.
    Benhar M, Dalyot I, Engelberg D, Levitzki A (2001) Enhanced ROS production in oncogenically transformed cells potentiates c-Jun N-terminal kinase and p38 mitogen-activated protein kinase activation and sensitization to genotoxic stress. Mol Cell Biol 21:6913–6926PubMedCrossRefGoogle Scholar
  30. 30.
    Saeki K, Kobayashi N, Inazawa Y et al (2002) Oxidation-triggered c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein (MAP) kinase pathways for apoptosis in human leukaemic cells stimulated by epigallocatechin-3-gallate (EGCG): a distinct pathway from those of chemically induced and receptor-mediated apoptosis. Biochem J 368:705–720PubMedCrossRefGoogle Scholar
  31. 31.
    Noguchi T, Ishii K, Fukutomi H et al (2008) Requirement of reactive oxygen species-dependent activation of ASK1-p38 MAPK pathway for extracellular ATP-induced apoptosis in macrophage. J Biol Chem 283:7657–7665PubMedCrossRefGoogle Scholar
  32. 32.
    Yamamori T, Inanami O, Sumimoto H et al (2002) Relationship between p38 mitogen-activated protein kinase and small GTPase Rac for the activation of NADPH oxidase in bovine neutrophils. Biochem Biophys Res Commun 293:1571–1578PubMedCrossRefGoogle Scholar
  33. 33.
    Brown GE, Stewart MQ, Bissonnette SA, Elia AE, Wilker E, Yaffe MB (2004) Distinct ligand-dependent roles for p38 MAPK in priming and activation of the neutrophil NADPH oxidase. J Biol Chem 279:27059–27068PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Joo Young Kim
    • 1
  • Su-Jin Yu
    • 1
  • Hyun Ju Oh
    • 1
  • Ji Young Lee
    • 1
  • Yongjin Kim
    • 1
  • Jeongwon Sohn
    • 1
  1. 1.Department of BiochemistryKorea University College of Medicine and Korean Institute of Molecular Medicine and NutritionSungbuk-GuKorea

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